<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0609</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2026-09-16</periodOfReport>

    <issuer>
        <issuerCik>0001641281</issuerCik>
        <issuerName>Bolt Biotherapeutics, Inc.</issuerName>
        <issuerTradingSymbol>BOLT</issuerTradingSymbol>
        <issuerForeignTradingSymbol></issuerForeignTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001724190</rptOwnerCik>
            <rptOwnerName>Quinn William P.</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerNonUSAddressFlag>false</rptOwnerNonUSAddressFlag>
            <rptOwnerStreet1>C/O BOLT BIOTHERAPEUTICS, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>900 CHESAPEAKE DRIVE</rptOwnerStreet2>
            <rptOwnerCity>REDWOOD CITY</rptOwnerCity>
            <rptOwnerState>CA</rptOwnerState>
            <rptOwnerZipCode>94063</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>true</isDirector>
            <isOfficer>true</isOfficer>
            <isTenPercentOwner>false</isTenPercentOwner>
            <isOther>false</isOther>
            <officerTitle>President, CEO and CFO</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>false</aff10b5One>

    <nonDerivativeTable>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-06-05</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>P</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
                <footnoteId id="F1"/>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>637</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.876</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>2950</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-16</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>1889</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.7740</value>
                    <footnoteId id="F2"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>41061</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-17</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>6545</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.3183</value>
                    <footnoteId id="F3"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>34516</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-18</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>9132</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.2364</value>
                    <footnoteId id="F4"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>25384</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">These shares were acquired under the Issuer's Employee Stock Purchase Plan in transactions that were exempt under Rule 16b-3(c).</footnote>
        <footnote id="F2">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.60 to $4.14. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
        <footnote id="F3">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.22 to $3.67. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
        <footnote id="F4">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.15 to $3.41. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ William P. Quinn</signatureName>
        <signatureDate>2026-09-21</signatureDate>
    </ownerSignature>
</ownershipDocument>
